Methods and compositions employing red rice fermentation products

ABSTRACT

Methods and compositions are disclosed which comprise red rice fermentation products, that can be used as natural dietary supplements and/or medicaments for the treatment or prevention of hyperlipidemia and associated disorders and symptoms, such as cardiovascular diseases, cerebrovascular diseases, diabetes, hypertension, obesity, asthenic breathing, chronic headache, chest pain and tightness, limb swelling and distention, loss of appetite and excess expectoration. The methods and compositions are effective in lowering both the serum cholesterol and serum triglyceride levels in humans, and can be used for maintaining cardiovascular health. The invention also encompasses particular Monascus strains that yield fermentation products with the desired biological activities.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation in part of U.S. patentapplication Ser. No. 08/720,548, filed Sep. 30, 1996, incorporatedherein by reference in full.

FIELD OF THE INVENTION

[0002] The invention relates to the fields of rice fermentation andtreatment of hyperlipidemia. More particularly, the invention relates tored rice fermentation products and methods, and use of the products totreat high cholesterol levels and other disorders.

BACKGROUND OF THE INVENTION

[0003] The invention relates to compositions comprising red ricefermentation products, that can be used as dietary supplements and/ortherapeutic medicaments. For example, the compositions can be used tolower serum cholesterol and triglycerides in mammals. Further, theinvention relates to methods of treating cardiovascular disorders andother diseases using the red rice fermentation products. In addition,the invention relates to particular Monascus strains that yieldfermentation products with the desired biological activities.

[0004] Red Rice in Ancient China

[0005] Red rice is known mostly for its use in food as a preservativeand colorant, and its uses in the dye industry. Red rice (known inChinese as Hung-ch'u or Hongqu) has also been known and used forhundreds of years in China in rice wine making and as a foodpreservative. In addition, red rice has been known as an ancient Chinesemedicine or an ingredient in certain ancient Chinese prescriptions.

[0006] Red Rice was first used around the time of the Han Dynasty. TaoGu, who lived in the age of Wudai after the Tang Dynasty, recorded “RedYeast Rice Cooked with Meat,” in Qing Yi Lu. The method of making RedRice was originally recorded in T'ien Kyng K'ai Wu and Pen Ts'ao KangMu, A detailed description of the medical applications of red rice wasprovided in the ancient Chinese pharmacopoeia, Pen Ts'ao Kang Mu, whichwas published during the Ming dynasty (1368-1644). In Pen Ts'ao Kang Mu,Red Rice is described as mild, nonpoisonous, and useful for treatingindigestion and diarrhea. Red Rice is also described as useful forimproving blood circulation and promoting the health of the spleen andstomach. Furthermore, several “prescriptions” using red rice fortreating aliments, such as indigestion, diarrhea, and heart andabdominal pains, are also provided in this ancient work. In accordancewith the Traditional Chinese Medicine Standard set forth inPharmacopoeia of People's Republic of China and the Traditional ChineseMedicine standard of Beijing, Nei Monggol, Shadog Provice, JiangsuProvince and Hunan province, etc., Red Rice is specified to be used as atraditional Chinese medicine. Furthermore, in the textbooks of Chineseuniversities and colleges such as Food Additives and Food Chemistry, RedRice is considered as additives for food and beverages, and has beenwidely used in the food processing industry for the production of suchitems as fermented bean curd, beer, and meat.

[0007] In an abbreviated English translation of Pen Ts'ao Kang Mupublished in 1911, red rice is described as useful for fermentation, andhaving medicinal value in the treatment of postpartum difficulties inwomen and dyspeptic conditions of children (Stuart, M. D., in “ChineseMateria Medica—Vegetable Kingdom,” page 233-234, republished in 1979 bySouthern Materials Center, Inc., Taipei, Republic of China). Red rice,as described in Pen Ts'ao Kang Mu, was subsequently recognized to be thefungal species known as Monascus purpureus Went (Read, B. E., 1936,Chinese Medicinal Plants from the Pen Ts'ao Kang Mu, 3rd edition,published by Peking National History Bulletin; Klein, G., 1932, Handbuchder Pflanzenanalyse II, p. 1422-1423, Wien, Verlag von Julius Springer).

[0008] The manufacture of red rice is taught in another publication fromthe Ming dynasty, T'ien Kung K'ai Wu by Sung Ying-Hsing, which waspublished in 1637 A.D. (see pages 291-294 in English translation of thisancient writing, “T'ien Kung K'ai Wu—Chinese technology in theseventeenth century,” translated by E-tu Zen Sun and Shiou-Chuan Sun.The Pennsylvania State University Press 1966). Red rice is describedtherein as useful for preserving the color and taste of fish or meat.The manufacturing process used red wine mash and cooked nonglutinousrice as starting materials. The method of making red rice by allowingthe fungus to grow on the surface of cooked rice was also recorded byVoderman (1894, Analecta ob Cromatologisch Gebied. II. Geneesh.Fylschrift voor Ned. Indie, 35, No.5).

[0009] Modernly, red rice, the fermentation product of Monascus species,is still used in traditional Chinese medicine, wine making and foodcoloring in Asia and Asian communities in North America. The red andyellow pigments of Monascus purpureus, such as monascorubin andmonascin, have been purified and extensively studied (Fielding et al.,1961, J Chem Soc, 4579-4589). The culture conditions and its effect onpigmentation of Monascus purpureus have also been studied (Broder etal., 1980, J Food Sci, 45:567-469). Antibacterial activity, especiallyagainst Bacillus species, was also detected in Monascus purpureusextract (Wong, 1977, Plant Physiol, 60:578-581). The Red Rice of thetraditional methods has been shown to be of little value and thus hasgradually fallen out of use in medical applications. The traditional RedRice has little effect of reducing blood lipids, and thus has never beenused as a cholesterol lowering agent.

[0010] Hyperlipidemia and Dietary/Medical Intervention

[0011] Lipids and lipoproteins play an essential role in transportingfat-derived metabolites between organs for absorption, metabolism, anddistribution (Felig et al., 1975, N Eng J Med, 293:1078-1084). Thesusceptibility to dietary-induced elevations in blood lipids includingcholesterol is extremely common. The interaction of geneticpredisposition and a high fat, high caloric diet coupled withunderactivity can lead to heart disease, hypertension,hypertriglyceridemia, and diabetes in a significant proportion of theUnited States population.

[0012] High serum cholesterol is a major risk factor for coronary arterydisease. Cholesterol is a major component of atherosclerotic plaque.Other associated lipid abnormalities, including hypertriglyceridemiaespecially in the presence of lowered HDL cholesterol levels, have beenrecognized as contributing to the risk of cardiovascular disease. Thereis a reciprocal relationship between elevated triglyceride levels andlowered HDL levels.

[0013] The level of cholesterol in circulation results from the balancebetween production of apoB-100 particles and its removal from thecirculation. Cholesterol is synthesized from acetyl-CoA via a series ofmore than 20 enzymatic reactions. This biosynthetic pathway is mainlyregulated by the activity of HMG-CoA reductase (hydroxymethylglutarylcoenzyme A reductase), which catalyzes the reduction of HMG-CoA tomevalonate. Since the majority of cholesterol circulating isendogenously synthesized in the liver, and not derived from dietarycholesterol, inhibitors of enzymes that are involved in the biosynthesisof cholesterol have been explored as drugs for the treatment ofhypercholesterolemia (Grundy, New Eng J Med (1988) 319:24-33).

[0014] One class of compounds inhibits cholesterol biosynthesis bycompeting with a natural substrate (HMG-CoA) for the key enzyme in thecholesterol biosynthetic pathway, HMG-CoA reductase. The first suchhypocholesterolemic compound discovered was compactin, which wasisolated from cultures of Penicillium citrinum by Akira Endo (Endo etal., J Antibiotics (1975) 29:1346-1348, see also U.S. Pat. Nos.3,983,140, 4,049,495, and 4,137,322). The hypocholesterolemic activityof this compound was demonstrated in several animal species (Tsujita etal., Atherosclerosis (1979) 32:307-313). Thereafter, ahypocholesterolemic compound structurally related to compactin wasindependently discovered by Endo in fermentation products of Monascusruber (the active compound was named monacolin K; Endo, J Antibiotics(1979) 32:852-854; Endo, J Antibiotics (1980) 33:334-336; see alsoGerman patents DE 3051175, 3051099 and 3006216; British patents GB2046737 and 2055100), and by another group from cultures of Aspergillusterreus. The active compound was also named mevinolin, lovastatin orMevacor®; Tobert et al., J Clin Invest (1982) 69:913-919), and has beenavailable in the United States since 1987 as a prescription drug. Theefficacy and long term adverse effect of this active compound has beenreviewed (Tobert, Am J Cardiol, 62:28J-34J). The isolated activecompound, its derivatives and methods of production from Aspergillushave been reported; see U.S. Pat. Nos. 4,231,938, 4,342,767, 4,294,926,4,319,039, 4,294,926, 4,294,846, and 4,420,491.

[0015] Although monacolin K or mevinolin has been successfully used totreat hypercholesterolemia, the compound has little or insignificanteffect on the serum level of triglycerides. Other lipid regulatingagents that have been used to treat hypertriglyceridemia, especiallytype IV and V hyperlipidemia, include nicotinic acid (e.g., niacin), andfibric acid derivatives (e.g., gemfibrozil and clofibrate). However, theuses of such agents are restricted because of their side effects, forexample, high doses of niacin may cause gastric irritability,hyperuricemia, hyperglycemia, pruritus, and gemfibrozil may lead tomalignancy, gall-bladder diseases, and abdominal pain. Moreover, therisk of myositis and rhabdomyolysis that can result in renal failureincreases when monacolin K is combined with gemfibrozil, clofibrate orniacin. Such combinations are only used with careful supervision inspecial situations that warrant the risk (The Merck Manual, 1992, 16thedition, pages 1044-1046). High concentrations of serum triglyceridesare known to be a risk factor for a variety of disease states and canlead to medical complications. Thus, there is a need for the developmentof a composition that accomplishes the reduction of the serum levels ofboth cholesterol as well as triglycerides. Regular exercise, propernutrition, and weight reduction programs can prevent or reduce theincidence of common chronic diseases such as heart disease associatedwith elevations of blood lipids (Pi-Sunyer, Am J Clin Nutr (1991)53:1595S-1603S). The role of diet in maintaining optimal health, and inslowing and reversing the progression of disease, has been the subjectof much research and public attention. The development of an effectivedietary supplement for use in the treatment of mixed hyperlipidemia,which could be used either with or without dietary changes, would be asignificant benefit.

SUMMARY OF THE INVENTION

[0016] The invention relates to a product of the fermentation of atleast one Monascus strain that can be used as a dietary supplement or asa therapeutic medicament to lower both serum cholesterol andtriglyceride levels in humans. The invention is based, in part, on thesurprising discovery that certain red rice products, i.e., the productof the fermentation of certain strains or mixtures of strains ofMonascus, are effective at lowering not only the level of serumcholesterol but also the level of serum triglyceride in mammals,particularly humans. Since monacolin K and mevinolin are not known to besignificantly effective in lowering serum triglyceride level, thebeneficial effect of red rice products is likely to be related to othercomponents in the fermentate.

[0017] In various embodiments of the invention, red rice can be used asa natural dietary supplement or a medicament to treat or prevent avariety of diseases, including but not limited to cardiovasculardiseases, diabetes, fatty liver conditions, stroke, cerebral thrombosis,hypotension, hypertension and obesity, and to modulate the circulatinglevels of lipids, such as cholesterol and triglyceride. In addition, thepresent invention encompasses methods for treating or preventing thesediseases in a human, which comprise administering to the human atherapeutically effective amount of a red rice fermentation product. Thepresent invention also encompasses methods for improving or maintainingcardiovascular health in a human comprising administering to aneffective amount of red rice fermentation product. The present inventionfurther encompasses methods for reducing the serum cholesterol and serumtriglyceride levels to normal levels in a human comprising administeringto the human a therapeutically effective amount of a red ricefermentation product. Red rice can also be used to treat or prevent avariety of ailments or symptoms as related to diseases of thecardiovascular system.

[0018] According to the invention, red rice can be manufactured invarious dosage forms and formulations. Also disclosed are methods formanufacturing red rice which are based on the traditional fermentationprocedures.

[0019] The terms “red rice fungi” or “Monascus” as used herein refer tothe prefermented organism, while the terms “red rice,” “red riceproduct”, “red rice extract” and the like refer to a product thatresults from the fermentation of at least one Monascus. Further, theselatter terms include traditional and improved red rice products asdescribed below. More specifically, “red rice product” as used hereinrefers to the product of fermentation, e.g., the fermentate of one or amixture of Monascus fungus. A “lovastatin-producing” Monascus strain(such as strain 0272) is one which can be fermented to produce a producthaving a lovastatin content of at least 0.05%, preferably at least 2%.

[0020] The red rice product is the fermentation product of at least oneof the following Monascus fungi set forth in the table below

[0021] Red rice is the fermentation product of one or a mixture ofMonascus fungi, comprising chiefly Monascus purpureus Went, and inlesser proportions other Monascus species, e.g., Monascus ruber vanTieghem, Monascus Fuliginosus Sato, Monascus Pilosus Sato and Monascusalbidus Sato. Red rice can also be the fermentation product of thefollowing strains of monascus fungi: Strains Accession No. Monascusalbidus Sato AS 3.570 AS 3.4440 CGMCC No. 0317 Monasuc pilosus Sato AS3.4444 AS 3.4633 AS 3.4646 AS 3.4647 Monascus pubigerus Sato AS 3.4445Monascus ruber van Tieghem AS 3.549 CGMCC No. 0315 CGMCC No. 0316Monascus paxii Lingelsheim AS 3.4453 Monascus fuliginosus Sato AS 3.569AS 3.1098 AS 3.2091 AS 3.2093 AS 3.2134 IFFI 05035 Monascus purpureusWent CGMCC No. 0272

[0022]Monascus purpureus Went ATCC 30141, AS 3.562, AS 3.991, AS 3.4446[ATCC 16365], AS 3.4642 [NRRL 2897], AS 3.4643 [NRRL 96], AS 3.4644, AS3.4645, AS 3.4651; Monascus ruber van Tieghem AS 3.549, IFFI 05007, IFFI05008, IFFI 05010, IPPI 05011; and Monascus anka IFFI 05038 (referencenumbers provided in China Catalogue of Cultures, 1992, China Committeefor Culture Collection of Microorganism, China Machine Press, Beijing1992). The improved red rice of the invention comprises Monascuspurpureus Went mutant strain M4027, 4028 and M4184.

[0023] The term “traditional red rice” as used herein refers to a redrice product which is the result of fermentation using a mixture ofMonascus fungi that has been used traditionally to manufacture red rice.“Traditional red rice” will generally contain less than about 0.005%lovastatin by weight. According to the invention, an “improved red rice”is produced by fermentation using one or more natural or mutant strainsof Monascus species, which yield a fermentate with improved biologicalor nutritional properties, e.g., higher hypocholesterolemic andhypotriglyceridemic activities than traditional red rice. The improvedred rice of the invention comprises Monascus purpureus Went mutantstrain CGMCC No. 0272. Several other strains can be used to achieve theobjectives of the invention. Improved red rice is sometimes referred toas Xuezhikang herein.

[0024] Generally, the red rice products of the present invention arered-purple powders that have a slightly bitter but mild and pleasanttaste. Similarly, the red rice products have a pleasant odor. The colorand/or odor may vary with the fermentation process, the strains used andthe processing steps. The red rice products of the invention contain atleast 0.05% lovastatin, more preferably at least about 2.0% lovastatinby weight.

[0025] As used herein, the term “effective treatment” means thereduction of a particular symptom, or the significant change of aparticular laboratory test toward the normal value. Preferably symptomsare relieved by at least 30-70% and a laboratory test is moved at least10% toward the normal value; more preferably symptoms are reduced by 70%and/or a laboratory test is moved at least 20% toward the normal value;most preferably, a treatment is effective if the symptoms are reduced by90%, and/or laboratory parameters are returned to the normal value.

[0026] The term “hypercholesterolemia” means the presence of elevatedlevels of cholesterol in the blood.

[0027] The term “therapeutically effective amount” or “therapeutic dose”as used herein means the amount of a particular agent sufficient toprovide a therapeutic benefit in the treatment or prevention of adisease, or in modulating the level of serum lipids and lipoproteins.

[0028] The term “dietary supplement” as used herein means an additionalelement that is added to the daily food intake of a mammal, usually ahuman.

[0029] Unless otherwise defined, all technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which this invention belongs. Although methods andmaterial similar or equivalent to those described herein can be used inthe practice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In case of conflict, the presentspecification, including definitions, will control. In addition, thematerials, methods, and examples are illustrative only and not intendedto be limiting.

[0030] Other features and advantages will be apparent from the followingdetailed description, and from the claims.

DETAILED DESCRIPTION OF THE INVENTION

[0031] The invention relates to compositions comprising the product ofthe fermentation of at least one Monascus species. These compositionsare useful for reducing the levels of both serum cholesterol and serumtriglycerides in mammals, and in particular humans. In addition, thecompositions are useful for modulating the levels of both serumcholesterol and triglycerides to maintain healthy levels despiteintrinsic (e.g. aging) or extrinsic (e.g. stress) factors that affectserum cholesterol and triglyceride levels. The compositions and methodsof the present invention are based, in part, on the discovery that thefermentate of Monascus species display hypocholesterolemic propertiesand also, unexpectedly, the ability to lower serum triglyceride levels.Since monacolin K is known not to be significantly effective in loweringserum triglyceride level, the beneficial effect of red rice productsmust be related to other components of the fermentate. The ability ofred rice products to lower serum triglyceride level provides the artwith a unique, natural alternative to the use of prescriptionhypocholesterolemic compounds.

[0032] According to the invention, traditional or improved red rice canbe prepared by traditional fermentation procedures or by modification ofthe traditional procedures. According to the earliest reported method(Sung, 1637, T'ien Kung K'ai Wu; pages 291-294, English translation bySun et al., Pennsylvania State Press 1966), red rice can be prepared bythe fermentation of washed and cooked nonglutinous rice using red winemash, natural juice of Polygonum grass, and alum water. The rice isfermented in open air for 7 days on bamboo trays under very cleanconditions. The rice changes its color from white to black, black tobrown, brown to red and then red to yellow, which is then harvested asred rice. According to an alternative traditional method, nonglutinousrice can be fermented in a hole in the ground lined by bamboo mats,which is securely covered. Fermentation is allowed to take placeunderground for one year or more, up to four years.

[0033] With respect to the present invention, the traditional method hasbeen improved by use of modern fermentation techniques and equipment tomore precisely control temperature, pH, pressure and other fermentationparameters, which, inter alia, reduces the time of fermentation. The keyfeature of the improved red rice preparation is that it contains activeingredients that can prevent or treat hyperlipidemia and relatedcardiovascular diseases. The preparations can be made as follows:

[0034] Preparation of Conventional Culture Fluid

[0035] For all of the media preparations rice or another grain is usedas a carbon source. The carbon source can be rice (polished long-grainnonglutinous rice, polished round-grain nonglutinous rice, polishedglutinous rice, red rice, and black rice), millet, barley, wheat, orcorn. Additionally sugar and substances containing sugar can be used.Organic compounds such as glycerine and glyceride can also be used inthe media preparations. For each 100 g of polished round-grainednonglutinous rice, 30-80 ml of culture medium are added. The culturemedia's key feature is that the carbon source is selected from the groupconsisting of cereals, sugar, and organic compounds, the source ofnitrogen is selected form the group consisting of beans (e.g. soya beanpowder, pressed soybean cake), or peanut powder (or pressed peanutcake), peptone, rice extract powder, thick beef juice, silkwormchrysalis powder, or inorganic salts (e.g. NH₄NO₃, etc.), and a sourceof phosphorous can also be added, such as inorganic salts (e.g. KH₂PO₄,K₂HPO₄, etc). Other inorganic salts can also be added, such as MgSO₄ orFeCl₂. By way of an example, and not by limitation, media preparationsof the invention are listed below:

[0036] Media 1: Liquid strain

[0037] 2-7% glycerine (or malt or potato juice)

[0038] 2-6% sugar

[0039] 0-3% peptone

[0040] 0.5-3% yeast extract powder

[0041] 0-3% thick beef juice (optional)

[0042] 2-4% defoamer (e.g. bean oil or peanut oil)

[0043] water

[0044] Media 2: Solid strain

[0045] 0-5% Potato juice

[0046] 0-6% sugar

[0047] 0-1.5% yeast extract or peptone

[0048] 30-80 ml of water per 100 g rice

[0049] Media 3:

[0050] 2-4% potato juice

[0051] 2-6% sugar

[0052] 0.5-3% yeast extract powder (or peptone or thick beef juice)

[0053] water.

[0054] Approximately 40-80 ml of the mixture is added to each 100 g ofrice, the pH is maintained at 3-8, and it is sterilized in steam at 121°C.

[0055] Generally, the pH is adjusted to 3.0-5.0, and the mixture issteam sterilized (121° C.). The mixture is cooled to 40° C., and therice is inoculated with the a Monascus strain of the invention. Forexample, the Monacus purpureus Went strain CGMCC No. 0272 is added andcultured at 15-35° C. for 9 days. Fermentation of the rice mixture ispreferably carried out at a temperature of 15-35° C., most preferably20-28° C., for over 4 days, most preferably 9 days or more, until theformation of Red Rice is noted. Any one of a number of methods offermentation, well known to one of skill in the art, can be used. Forexample, an Erlenmeyer flask, tray, or ventilated fermentation bed canbe used as fermentation facilities. At the end of the fermentationprocess, the fermentation broth is drained and discarded, while thesolid residue is sterilized by heat (for example, by high pressuresteam). For example, the fermentation product is sterilized at atemperature of 69-121° C., and dried. This dried product can be ground.Standard mesh sizes for the production of capsules, tablets, powders andsuspensions are well known in the art. By way of example, the improvedred rice of the invention can be ground to 80 mesh under vacuum at atemperature of approximately 60-80° C., and the powdered productrecovered. This product can be used directly in the various compositionsand formulations provided by the present invention. For example, it canbe filled into capsules. Alternatively the 80 mesh ground product canfurther be ground to 200 mesh. The 200 mesh powder can then formulatedinto tablets using standard methodologies. Alternatively, liquid orsyrup formulations of red rice can be made using conventionalprocedures.

[0056] Optionally, the dried crushed red rice powder can be furtherprocessed, e.g., extracted with organic solvents, such as but notlimited to, alcohols (e.g. 75-90% ethanol) to remove starch and/or agar.After evaporation to dryness, the extract can be used in the variouscompositions and formulations as provided by the present invention. Theextracted product can further be concentrated under a vacuum andevaporated (60-80° C., 0.06-0.08 MPa) until dry. This provides anexceptionally useful supplement at very low cost.

[0057] According to the invention, an “improved red rice” is produced byfermentation using one or more natural or mutant strains of Monascusspecies, which yield a fermentate with improved biological ornutritional properties, e.g., higher hypocholesterolemic andhypotriglyceridemic activities than traditional red rice. The improvedred rice of the invention comprises Monacus purpureus Went mutant strainM4027, 4028 and M4184. Several other strains can be used to achieve theobjectives of the invention (Chinese Microorganism Strain Index, 1992,China Microorganism Collection Committee), as listed in the Table above.

[0058] Lovastatin in red rice may be extracted using 10 ml of 75% EtOHat ambient temperature. The extract (2 ml) is treated with 1 ml 0.06 MNaOH in 75% EtOH for 30 min, then with 1 ml 0.06 N H₃PO₄ (in 75% EtOH),and the mixture applied to a C18 HPLC column (150×4.60 mm), anddeveloped with 0.02 N H₃PO₄ (in 70% MeOH) to quantify the amount oflovastatin present.

[0059] Red rice can be used as a natural dietary supplement or apharmaceutical medicament to prevent illness (maintain health) or totreat or a variety of diseases, including but not limited tocardiovascular diseases, diabetes, stroke, hypertension and obesity, andto modulate the circulating levels of lipids and lipoproteins, such ascholesterol and triglycerides. Red rice can also be used to treat orprevent a variety of symptoms related to these above-mentioned diseasesand associated with poor cardiovascular health due to aging and otherintrinsic and extrinsic factors.

[0060] As used herein, examples of cardiovascular diseases may includebut are not limited to myocardial infarction, coronary heart disease,atherosclerosis, arteriosclerosis. The present invention includes thetreatment or prevention of cerebrovascular disease such as stroke,memory loss due to stroke, and cerebral thrombosis.

[0061] The present invention also encompasses a composition comprising atherapeutically effective amount of a red rice product, for example 2-4grams per day, useful in humans for the treatment or prevention ofhyperlipidemic disease, cardiovascular disease, cerebrovascular disease,hypertension, hypotension, diabetes, fatty liver conditions, or obesity,or a combination thereof.

[0062] The present invention further encompasses a compositioncomprising a therapeutically effective amount of a red rice product,useful for the modulation of serum lipid and lipoprotein levels in ahuman in need of therapy to maintain the lipid and lipoprotein levelswithin a healthy normal range. In one embodiment of the invention, thecomposition is adapted for use in the treatment or prevention ofhypertriglyceridemia. In a preferred embodiment, such a composition isused for reducing serum cholesterol and serum triglyceride levels inhumans.

[0063] The present invention further encompasses a compositioncomprising a therapeutically effective amount of improved red riceproduct, useful for the treatment of any one of the following symptoms:shortness of breath; asthenic breathing; lethargy; dizziness; chronicheadache; chest pain and tightness; heartache; loss of appetite; limbswelling; tightness and distention.

[0064] Functions of Improved Red Rice

[0065] The improved red rice of the invention and preparations of theimproved red rice contain statinoid compounds, i.e. hydroxy-acidLovastatin and lactone Lovastatin.

[0066] Without being bound by theory, the above-mentioned Monascusstrains, when cultured under the appropriate fermentation conditions,have an increased content of the statinoid compounds. The increase inthe statinoid compounds reduces serum cholesterol and serumtriglycerides, while increasing high-density lipoprotein cholesterolsimultaneously.

[0067] The red rice of the invention can be employed to treathyperlipidemia and other related cardio-cerebrovascular diseases, suchas atherosclerosis, coronary heart disease, myocardial infarction,diabetes, hypertension, and cerebral embolism, among others.

[0068] Method of Treatment

[0069] The present invention provides methods for treating a humanafflicted by a variety of diseases, disorders, and symptoms. In additionto treatment of a human disease, the methods of the invention can alsobe used for preventive treatment in a person susceptible to suchdiseases, disorders or symptoms.

[0070] The invention encompasses methods of treatment of hyperlipidermicdisease, cardiovascular disease, cerebrovascular disease, hypertension(hereditary and non-hereditary), hypotension, angina, stroke, diabetes,fatty liver conditions, or obesity, or a combination thereof in a human,comprising administering to the human a therapeutically effective amountof a red rice product, or compositions containing said product.

[0071] The invention also encompasses methods of preventinghyperlipidemic disease, cardiovascular disease, cerebrovascular disease,hypertension, hypotension, angina, stroke, diabetes, fatty liverconditions such as fatty liver deposits, obesity or a combinationthereof, which comprises administering an effective amount of a red riceproduct of the present invention. The method of the invention ispreferably used to treat or prevent hypertriglyceridemia and associateddiseases, such as diabetes, in humans.

[0072] As used herein, examples of cardiovascular diseases may includemyocardial infarction, coronary heart disease, atherosclerosis,arteriosclerosis, and cerebrovascular diseases or conditions, includingstroke, cerebral thrombosis or memory loss due to stroke.

[0073] The present invention also provides methods for modulating serumlipid and lipoprotein levels in a human in need of lowering the lipidand lipoprotein levels to a healthy normal range, which compriseadministering to the human a therapeutically effective amount of a redrice product, or compositions containing said product. In a preferredembodiment, the method of the invention is used to reduce serumcholesterol and serum triglyceride levels in a human. The methods of theinvention are particularly useful for the treatment of geriatricpatients and postmenopausal women.

[0074] The present invention further provides methods for treating ahuman afflicted by shortness of breath, asthenic breathing, lethargy,dizziness, chronic headache, loss of appetite, limb swelling, tightnessand distention, and abdominal distention, or a combination thereof,which comprises administering to the human a therapeutically effectiveamount of a red rice product, or compositions containing said product.

[0075] The preventive or therapeutic dose of traditional red rice orimproved red rice in the treatment or prevention of diseases and in themanagement of serum lipid and lipoprotein levels will vary with thecondition to be treated and the severity of the condition to be treated.The dose, and perhaps the dose frequency, will also vary according tothe age, body weight, and response of the individual patient. Ingeneral, the total daily dose range of red rice, for the conditionsdescribed herein, is from about 0.1 g to about 5 g administered insingle or divided doses orally. For examples a preferred oral daily doserange should be from about 0.3 g to about 4 g, while most preferably anoral daily dose should be about 1.2 to about 2.5 g. For example, twocapsules each containing 0.6 g of red rice may be taken orally twice aday to obtain the preferred dose. A course of treatment should be atleast 4 weeks. It may be necessary to use dosages outside these rangesin some cases as will be apparent to those skilled in the art. Further,it is noted that the nutritionist, dietitian, clinician or treatingphysician will know how and when to interrupt, adjust, or terminatetherapy in conjunction with individual patient response.

[0076] It should be noted that the present invention encompasses newuses of traditional red rice, and novel red rice products and novelmethods of using those products.

[0077] Dietary Supplement Use

[0078] As mentioned above, the present invention encompassescompositions and methods of using traditional and novel or improved redrice products as dietary supplements. As such, the red rice productsprovide the individual with a means for maintaining normal or healthylevels of serum cholesterol and triglycerides despite intrinsicdeterioration, e.g., from aging and extrinsic factors such as stress,lack or exercise and poor nutrition. The dietary supplements alsoprovide means for preventing, or reducing the likelihood orexperiencing, the diseases discussed above. Finally, the dietarysupplements can be used to prevent weight gain or obesity. Finally, thedietary supplements containing red rice products are particularly usefulfor the elderly and postmenopausal women. The dietary supplements shouldbe taken daily for at least four weeks and can be used permanently on adaily basis. A daily dose is from about 0.1 g to about 5.0 g; preferablyabout 1 to about 4 g; and most preferably about 1.2 to about 2.4 gramsper day.

[0079] Formulation

[0080] The pharmaceutical and dietary compositions of the presentinvention comprise a red rice product, or an extract thereof, as activeingredient, and may also contain a pharmaceutically acceptable carrieror excipient and, optionally, other ingredients.

[0081] Other ingredients that can be incorporated into the dietary orpharmaceutical compositions of the present invention may include, butare not limited to, vitamins, amino acids, metal salts and flavorenhancers. For oral administration the compositions comprising red ricecan be added directly to foods so that a therapeutically effectiveamount of red rice is ingested during normal meals. Any methods knows tothose skilled in the art may be used to add or incorporate red rice tonatural or processed foods.

[0082] Compositions of the present invention suitable for oraladministration may be presented as discrete units such as capsules,cachets, or tablets, each containing a predetermined amount of a redrice product, as a powder or granules, or as a solution or a suspensionin an aqueous liquid, a nonaqueous liquid, an oil-in-water emulsion, ora water-in-oil liquid emulsion. In general, the compositions areprepared by uniformly and intimately admixing the active ingredient withliquid carriers or finely divided solid carriers or both, and then, ifnecessary, shaping the product into the desired presentation.

[0083] The compositions of the present invention may additionallyinclude binding agents (e.g., pregelatinized maize starch,polyvinylpyrrolidone or hydroxypropyl methylcellulose); binders orfillers (e.g., lactose, pentosan, microcrystalline cellulose or calciumhydrogen phosphate); lubricants (e.g., magnesium stearate, talc orsilica); disintegrants (e.g., potato starch or sodium starch glycolate);or wetting agents (e.g., sodium lauryl sulphate). The tablets orcapsules can be coated by methods well known in the art.

[0084] Liquid preparations for oral administration can take the form of,for example, solutions, syrups or suspensions, or they can be presentedas a dry product for constitution with water or other suitable vehiclebefore use. Such liquid preparations can be prepared by conventionalmeans with pharmaceutically acceptable additives such as suspendingagents (e.g., sorbitol syrup, cellulose derivatives or hydrogenatededible fats), emulsifying agents (e.g., lecithin or acacia), nonaqueousvehicles (e.g., almond oil, oily esters, ethyl alcohol or fractionatedvegetable oils), and preservatives (e.g., methyl orpropyl-p-hydroxybenzoates or sorbic acid). The preparations can also bemade to resemble foods, containing buffer salts, flavoring, coloring andsweetening agents as appropriate.

[0085] Any dosage form may be employed for providing the patient with aneffective dosage of the red rice product. Dosage forms include tablets,capsules, dispersions, suspensions, solutions, capsules and the like.Because of their ease of administration, tablets and capsules representthe most advantageous oral dosage unit form, in which case solidpharmaceutical carriers as described above are employed. In addition tothe common dosage forms set out above, the compounds of the presentinvention may also be administered by controlled release means. However,the most preferred oral solid preparations are capsules.

[0086] For example, a tablet may be prepared by compression or molding,optionally, with one more accessory ingredients. Compressed tablets maybe prepared by compressing in a suitable machine red rice in afree-flowing form such as powder or granules, optionally mixed with abinder, lubricant, inert diluent, surface active or dispersing agent.Most preferably, the composition is a capsule containing 0.3 g of redrice in powder form.

[0087] The invention is further defined by reference to the followingexamples describing in detail the human clinical trials conducted tostudy the efficacy and safety of red rice. It will be apparent to thoseskilled in the art that many modifications, both to materials andmethods, may be practiced which are within the scope of this invention.

[0088] The invention will be further described in the followingexamples, which do are intended to provide further description of theinvention, and are not intended to limit the scope of the claim.

EXAMPLE 1 Preparation of Red Rice Cultures

[0089] (A) A liquid strain culture fluid was prepared containing 2-4%glucose, 3-5% glycerine, 0-3% thick beef juice, 0.8-1.6% peptone, 0-3%yeast extract powder, 0.1% KH₂PO₄, and 0.05% MgSO₄.7H₂O and water. Twomedia were prepared, the first containing 2% glucose, 3% glycerine, 1.5%thick beef juice, 0.8% peptone, 3% yeast extract powder, 0.1% KH₂PO₄,and 0.05% MgSO₄.7H₂O in water, and the second containing 4% malt juiceor potato juice, 8% sugar, 1.5% yeast extract powder, 3% thick beefjuice, and water. The pH was adjusted to 3.5 using acetic acid. For each50 ml of culture fluid, 100 g of polished round-grained nonglutinousrice was added, and the media were sterilized in steam at 121° C. Themixture was cooled to below 40° C., and inoculated with Monascuspurpureus Went (CGMCC No. 0272) in glass tubes or plates and cultured at30-34° C. for 24-36 hours. Fermentation was continued at 25° C. for 9days. Once the fermentation period was completed, the mixture wassterilized at high temperature (100-121° C.), dried under a vacuum at80° C., and ground to 60-100 mesh. The powder was then filled intocapsules. Yield approximately 65%.

[0090] (B) Alternatively, culture fluid containing 4% malt juice orpotato juice, 8% glucose, 3% thick beef juice, 3% peptone and water (pHvalue adjusted to 3) was used. Following fermentation, the resultingproduct was extracted using 75-95% ethanol, the red rice was dried andfully mixed with untreated rice. Soya bean powder (10 g) and culturefluid (50 ml) were added to each 100 g mixture, and the composition wassterilized. The mixture was cooled to 30-40° C. and then inoculated with10-20 ml liquid Monascus strain (CGMCC No. 0272), fermented at 30-34° C.for 3-4 days and at 23-25° C. for over 15 days. The mixture wassterilized at 100-121° C. and dried under vacuum. This red rice productwas ground and tablets were produced.

[0091] (C) Another medium was prepared using 4% malt juice, 6% maltsugar, 1% yeast extract powder, 6% peptone and water (pH=3). Soya-beanpowder (15 g) and culture fluid (50 ml) was added to each 100 g ofpolished round-grained nonglutinous rice. The mixture was sterilized at121° C. and then inoculated with Monascus ruber AS 3.549 (20 ml).Fermentation was carried out at 25° C. for over 9 days. Aftersterilization, the mixture was dried at 80° C.

[0092] The Red Rice was further processed into concentrates usingalcohol (75%) extraction twice. After evaporation under vacuum, theconcentrated substance was obtained and the alcohol was recovered. Theresulting concentrated substance contained approximately 25 mglovastatin per gram red rice, and was used as raw material for theproduction of the capsules or tablets.

[0093] (D) Another culture fluid was prepared containing 4% malt juice,approximately 8% sugar, 2% yeast extract powder, 5% thick beef juice andwater (pH=3). Culture fluid (50 ml) was added to each 100 g of rice andthe mixture was sterilized in steam at 121° C. After cooling below 40°C., the mixture was inoculated with Monascus albidus (CGMCC No. 0317)and then fermented at 25° C. for over 12 days. The mixture wassterilized and then dried. This Red Rice preparation was ground to 200mesh and granulated with alcohol for pill preparation.

[0094] (E) Culture fluid was also prepared using 3% potato juice,approximately 6% sugar, 1.5% yeast extract powder, 4% peptone and water(pH=3). Soya bean powder (10 g) and culture fluid (80 ml) were added toeach 100 g of rice and the mixture was sterilized in steam at 121° C.After cooling below 40° C., the mixture was inoculated with Monascuspilosus Sato (AS 3.4444), then cultured at 30-34° C. for 3-4 days, thencultured for over 10 days at 100-121° C. and then dried.

[0095] This Red Rice preparation was further processed into concentratesby using alcohol (75%) extraction. Small amounts of dissolvable starchwere added for pill preparation.

[0096] (F) Culture fluid was also prepared using 3% potato juice, 5%sugar, 6% thick beef juice and water (pH=3). 10-20% peanut powder wasadded to culture fluid (80 ml) for each 100 g of rice, and the mixturewas sterilized in steam at 121° C. After cooling below 40° C., themixture was inoculated with Monascus ruber van Tiegheim (CGMCC No. 0315)and then cultured at 30-34° C. for 3 days. The temperature was loweredto 24° C. and the culturing was continued for over 15 days. The mixturewas sterilized in steam at 100-121° C. and dried.

[0097] (G) A medium was also prepared containing 3% corn juice or 3%potato juice, 6% sugar, 1.5% yeast extract powder, 4.5% peptone andwater (pH=3). 5-20% of soya-bean cake powder and 80 ml of culture fluidwere added to each 100 g of millet. The mixture was sterilized in steamat 121° C., cooled to a temperature below 40° C., inoculated withMonascus pilosus (AS 3.4633), and cultured at 25° C. for over 18 days.The mixture was sterilized at 121° C. and then dried under a vacuum at atemperature of 60-80° C.

[0098] (H) A medium was prepared containing 4% potato juice, 7% sugar,8% peptone and water (pH=3). Culture fluid (60 ml) was added to each 100g of rice, and the mixture was sterilized at 121° C. After cooling to atemperature below 40° C., the mixture was inoculated with Monascuspubigerus Sato (AS 3.4445), and then cultured at 30-34° C. for 3 days.The temperature was lowered to 25° C. and culturing was continued forover 9 days. The mixture was sterilized in steam at 121° C. and dried at80° C.

[0099] (I) A medium was prepared containing 5% soya bean milk, 5%glucose, 2% yeast extract powder, 5% soya-bean peptone and water (pH=3).Culture fluid (80 ml) was added to each 100 g of rice and the mixturewas sterilized in steam at 121° C. After cooling to a temperature below40° C., the mixture was inoculated with Monascus pilosus Sato (AS3.4646), and cultured at 30-34° C. for 3 days. The temperature waslowered to 23-25° C., and cultured for over 9 days. The mixture wassterilized in steam at 121° C. and then dried at 80° C.

[0100] (J) A medium was prepared containing 4% potato juice, 4% sugar,3% yeast extract powder and water (pH=3). Silkworm chrysalis powder (5g) and culture fluid (60 ml) were added to each 100 g of rice and themixture was sterilized in steam at 121° C. After cooling, to atemperature below 40° C., the mixture was inoculated with Monascusfuliginosus Sato (AS 3.569), and then cultured at 30-34° C. for 3 days.The temperature was lowered to 23-25° C. and culturing was continued forover 9 days. The mixture was sterilized in steam at 121° C. and driedunder vacuum at 60-80° C.

[0101] (K) A medium was prepared containing 3% malt juice, 5% sugar, 6%thick beef juice and water (pH=3). Culture fluid (80 ml) was added toeach 100 g of rice, and the mixture was sterilized in steam at 121° C.After cooling to a temperature below 40° C., the mixture was inoculatedwith Monascus fuliginosus Sato (AS 3.1098), and then cultured at 30-34°C. for 3 days. The temperature was lowered to 23-25° C. and culturingcontinued for over 9 days. The mixture was sterilized in steam at 121°C. and dried at a temperature of 80° C.

[0102] (L) The following procedures were used for large scalefermentation:

[0103] (1) Soaking the rice: Rice (500 kg) was placed in several layersof baskets. The chaff was cleaned in water, and the rice soaked in waterfor 16-24 hours. The rice was dredged from the water and dried (thecontent of water is approximately 22-24%).

[0104] (2) Steaming the rice: Dried rice was poured into a rice steamerand steamed for 50-70 minutes. The steamed rice was spread out on abamboo mat or in baskets, dispersed, and cooled to a temperature below40° C. The rice was then inoculated with approximately 20 kg of solidMonascus strain and 2.5-3 kg of acetic acid and stirred.

[0105] (3) Fermentation: For the first 3 days, the rice was turned overseveral times per day. The temperature was controlled between 30° C. and34° C. After 3 days, the temperature was reduced to 23-25° C. The ricewas turned over once daily, during which water (pH value adjusted to 3.5using acetic acid) was added at quantity depending on the humidity ofthe fermenting mixture. The mixture was fermented for over 15 days.

[0106] (4) Preservation: After the fermentation process, the mixture wassterilized, dried and preserved.

[0107] The large-scale fermentation methods were used with all media andpreparative processes described above by adding in the respectiveproportions of other ingredients. It should be noted that forcedventilation can be used in the fermentation process but that the returnair must be sterile.

EXAMPLE 2 Pharmacology and Toxicology

[0108] Pharmacological and toxicological studies of the red rice of thepresent invention were performed in experimental animal models. Red ricewas shown to dramatically decrease serum total cholesterol (TC) ofendogenous hyperlipidemic rabbits, remarkably decrease TC and totaltriglyceride (TG) of exogenous hyperlipidemic rabbits, inhibit formationof arteriosclerosis plaque and lipid deposition in liver inhyperlipidemic rabbits, and decrease serum TC and TG of hyperlipidemicquails.

[0109] In acute toxicity studies, a LD₅₀ value cannot be determined. Thehighest tolerance dose of red rice in mice is over 16 g/kg, which is 533times over the dose used in clinical treatment. Moreover, in otherexperiments, rats were continuously force-fed red rice for four months;no rats died or showed toxic symptoms due to this drug. Hematologicalindices, main viscera indices, blood biological indices, routineuroscopy and pathological examination did not show any differencesbetween experimental groups and control groups.

EXAMPLE 3 Human Clinical Study I

[0110] The following two examples contain the methodologies and resultsof two human clinical trials that were carried out in China. The trialswere designed to determine the efficacy of a red rice product inmodulating circulating serum lipid and lipoprotein levels in humans, inresolving symptoms according to traditional Chinese medicine, and inestablishing the safety of a red rice product.

[0111] In the first randomized human clinical trial, 446 patients withhyperlipidemia, who were also diagnosed as suffering from hypofunctionand disorder of the spleen by traditional Chinese medicine, were dividedinto two treatment groups.

[0112] The first group (324 patients) received Xuezhikang capsules,which contained 0.3 g of a red rice product. The second group (122patients) served as a control; they received Jiaogulan tabletscontaining a lipid-regulating drug (gynostemma pentaphyllum) that isbased on traditional Chinese herbal medicine.

[0113] All the patients with primary hyperlipidemia stopped using serumlipid modulators two to four weeks prior to the beginning of the trialand received dietary advice. Serum samples was taken and laboratorytests was conducted to determine eligibility for the study. Onlypatients who met the following criteria were enrolled in the trial:total serum cholesterol (TC)>230 mg/dl (5.95 mmol/L) and triglyceride(TG)>200 mg/dl (>2.26 mmol/L/L). High density lipoprotein cholesterol(HDL-C) was also considered as a reference; male<40 mg/dl (1.04 mmol/L),female<45 mg/dl (1.16 mmol/L). All patients were diagnosed as deficientin the function of the spleen and having excess expectoration bytraditional Chinese medicine. The patients also had the followingsymptoms: limb weakness; asthenic breathing; pain and oppressed feelingin chest; loss of appetite; distention and swelling on gastric region;whitish or purple dots on the tongue; thick-white or thick-slimy fur onthe tongue; taut-slippery or hesitant-weak pulse.

[0114] Patients who had the following disorder or disease were excludedfrom the trial: myocardial infarction; cerebrovascular disease; severewound or major surgery during the past half year; nephritic syndrome;hypothyroidism; acute and/or chronic hepatobiliary disorder; diabetes;gout; general allergic reactions; and psychosis.

[0115] The total number of patients enrolled was 446. In the grouptreated with red rice, there were 188 male and 126 female patients. Theratio of male versus female was 1.38:1 and the average age was 56.0±9years old. There were 73 male and 45 female patients in the controlgroup. The ratio of male versus female was 1.49:1 and average age was56.4±9.1 years old. Between the two groups, there was no difference(P>0.05) found in baseline parameters including age, sex and course ofdisease, serum lipid and lipoprotein levels.

[0116] The group receiving red rice treatment took two Xuezhikangcapsules orally, twice a day for 8 weeks. The control group took threeJiaogulan tablets twice a day for 8 weeks. All the patients maintainedthe same lifestyle and habits as before.

[0117] The following tests were performed at four and eight weeks:measurements of weight, blood pressure, and cardiac rhythm wereperformed. In addition, an electrocardiogram and routine physicalexamination was performed. The following parameters were monitored bylaboratory tests: blood urea nitrogen (BLTN); creatinine; serum glutamicpyruvic transaminase (SGPT, ALT); serum glucose; and creatinine kinase(CK).

[0118] To determine serum lipid and lipoprotein levels, a fasting (12hours) venous blood sample was taken from patients who were told not toconsume alcoholic beverages or food with a high fat content at the lastmeal prior to the tests. Serum obtained from the patients was separatedimmediately, and stored in −20° C. for analysis. TC, TG and HDL-C wereanalyzed, and the LDL-C value was calculated according to the formula:LDL-C=TC−HDL-C−(TG/2.2).

[0119] Efficacy of treatment was evaluated according to the criteria setforth in “Clinical trial management: hyperlipidemia treatment using newChinese materia medica” released by the Ministry of Health of China asfollows:

[0120] 1. Cure: All symptoms were eliminated or a reduction of the totalsymptom score by more than 90%, and a return of all laboratory testparameters to normal.

[0121] 2. Effective: Symptoms were significantly relieved, i.e., symptomscore reduced by 70%-89%. Serum lipid and lipoprotein did not reachnormal, but were improved in one of the following respects: 1) reducingTC≧20%; 2) reducing TG≧40%; 3) reducing (TC−HDL-C)/HDL-C≧20%; 4)increasing HDL-C>10 mg/dl.

[0122] 3. Improvement: symptoms were relieved, i.e., symptom scorereduced by 30%-69%. Serum lipid and lipoprotein levels were not normalbut were improved in one of the following respects: 1) reducing TC at10%-20%; 2) reducing TG≧20% but <40%; 3) reducing (TC−HDL-C)/HDL-C≧10%but <20%; 4) increasing HDL-C>4 mg/dl (0.14 mmol/L but <10 mg/dl.

[0123] 4. Inefficacy: Symptom score was reduced by less than 30%, andthe laboratory test parameters did not meet the criteria ofeffectiveness.

[0124] All the data are subjected to statistical analyses (the Student'st test, Chi-square test, Ridit assay for data of stratum, and U chartfor percentiles analysis were used as appropriate). TABLE 1 Efficacycomparison Case Cure Effective Improvement Inefficacy Total Improvementnumber n % n % n % n % Effective n % Treated 324 169 52.2 89 27.5 4413.5 22 6.8 258 79.7 302 93.2 Group Control 122 13 10.7 25 20.5 24 19.760 49.2 38 31.1 62 50.8 Group

[0125] TABLE 2 Comparison of serum lipid and lipoprotein levels aftertreatment Difference Difference after 4 after 8 Mean ± S week week CsseBaseline difference % difference % Parameter Group No. mg/dl in mg/dlChange in mg/dl Change TC Treated 251 273.5 ± 31.3 −47.4 −17.3** −62.8−23** Control 94 268.2 ± 25.4 −13.2 −4.9** −18.9 −7** TG Treated 183296.0 ± 75.5 −66.3 −22.4** −108 −36.5** Control 72   289 ± 71.7 −27.5−9.5* −42.3 −14.6** HDL-C Treated 121 35.9 ± 4.4 4.2 11.8** 7 19.6**Control 55 35.1 ± 4.0 1.8 5* 3 8.6** LDL-C Treated 324 162.2 ± 52.4−36.5 −22.5** −46.3 −28.5** Control 122 157.3 ± 49.2 −9 −5.7** −12.6−8** TC-HDL- Treated 324  4.69 ± 1.44 1.3 −22.7** 01.6 −34.2** C/HDL-CControl 122  4.79 ± 1.71 0.39 −8.1** −0.52 −10.9**

[0126] Table 1 shows a comparison of overall efficacy. The score for thegroup which received red rice (treated group) was much higher than thatin the control group (X²=9.7, P<0.001).

[0127] The percentage of patients in the treated group who reported theelimination of symptoms diagnosed by traditional Chinese medicine wasmuch higher than that in the control group (p<0.05-0.001). Thosesymptoms were: condition of tongue (whitish or purple dots on thetongue; thick-slimy fur); pulse (slippery-taut or hesitant-weak);oppressed feeling in chest; loss of appetite; abdominal distention andswelling.

[0128] With respect to serum lipid and lipoprotein level, the efficacyscores for curing or reducing total serum cholesterol and totaltriglyceride level in the treated group were greater than that in thecontrol group. The score for normalizing or increasing HDL-C, level andthe score for reducing Atherosclerotic Index in the treated group werealso much better than the control (P<0.001).

[0129] Table 2 indicates that both Xuezhikang-treated and control groupsshowed marked desirable changes in the levels of TC, TG,(TC−HDL-C)/HDL-C, HDL-C serum levels markedly. The effectiveness ofXuezhikang was found to be superior to that of Jiaogulan.

[0130] It was also observed that the higher the baseline of TC and TG inthe serum, the more effective is the reduction of TC and TG after usingXuezhikang. As for HDL-C level, a greater increase was observed aftertreatment in patients who had a lower starting baseline. TABLE 3 Effectsof Xuezhikang capsule on patients with different abnormal levels serumlipid and lipoprotein. TC (mg/g) TG (mg/g) HDL-C (mg/dl) Parameter <230230-300 >300 <230 230-300 >300 >45 35-45 <35 Case No. 73 206 45 141 11271 161 114 49 Mean 187.8 261.8 327.1 134.3 247.6 327.3 56.4 40.1 5.4baseline (mean) Difference ↓20.5 ↑42.5 ↓69.8 ↑2.7 ↓51.4 ↓89.8 ↑1.3 ↑4↑5.4 (4 weeks) % changes ↓10.9 ↓16.2 ↓21.3 ↑2 ↓20.8 ↓24.1 ↑2.3 ↑10 ↑17Difference ↓30.6 ↓57.9 ↓86.1 ↓15.9 ↓81.4 ↓149.9 ↑2.1 ↑6.3 ↑7.2 (8 weeks)% Changes ↓16.3 ↓22.1 ↓26.3 ↓11.8 ↓32.9 ↓40.2 ↑3.7 ↑15.7 ↑22.8Comparison ** ** ** ** * *

[0131] TABLE 4 Effect of Xuezhikang capsule on apoA-1 and apoB (mean ±S) Group Case No. time Point apoA-1 apoB apoB Treated 88 Baseline 1.22 ±0.19  1.2 ± 0.19 1.05 ± 0.25 Group 4 Weeks 1.32 ± 0.13(4) 1.09 ± 0.21(3)1.25 ± 0.27(5) ↑8.2% ↓9.2% ↑19% 8 Weeks 1.28 ± 0.13 0.99 ± 0.18(3) 1.33± 0.30(3) ↑4.9 ↓18% ↑26.7% Comparison 30 Baseline 1.19 ± 0.16 1.21 ±0.15 1.00 ±0.18 Group 4 Weeks 1.26 ± 0.11(1) 1.15 ± 0.17(1) 1.11 ±0.14(2) ↑5.9% ↓5% ↑11.0% 8 Weeks 1.26 ± 0.09(l) 1.03 ± 0.15(3) 1.24 ±0.21(3) ↑5.9% ↓14.9% ↑24.0%

[0132] Regarding the effect of Xuezikang on apolipoprotein as a-I(apoA-I) and apolipoprotein B (apoB), the serum levels of apoA-I in bothgroups were raised after therapy. Statistical results show a significantdifference in apoA-I levels after a four week treatment. ApoB levelswere reduced somewhat in both groups, however, these reductions are notstatistically significant. The treated group showed better improvementof apolipoprotein B and apoA-I/apoB over the control.

[0133] With respect to rheology, there were significant changes to inblood sedimentation and K-value in both groups after treatment(P<0.05-0.0 1). However, the treated group showed better results thanthe control group (P<0.05-0.0 1).

[0134] All 446 patients were subjected to the following laboratory testsbefore and after therapy: blood urea nitrogen (BUN); creatinine; serumglutaliic pyruvic transaminase (SGPT, ALT); serum glucose; andcreatinine kinase (CK); and routine examination of blood and urine. Noclinically meaningful changes were found at the end of the trial.

[0135] Several patients developed a burning sensation in the stomach(six patients, 1.8%), experienced fullness in the stomach (threepatients, 0.9%), and suffered dizziness (one patient 0.3%). All patientshad previously finished the trial, and all the symptoms werespontaneously relieved without treatment. Two patients sufferedgastritis after taking Xuezhikang and had to leave the trial. Theresults suggest that Xuezhikang is a safe and effective drug forlowering serum lipids and triglycerides.

[0136] In this trial, a lipid regulating agent known in traditionalChinese medicine was used as a positive control. The efficacy score inthe Xuezhikang-treated group was much higher than that in the controlgroup (P<0.001). Comparing the baseline, in Xuezhikang-treated group,serum level of high density lipoprotein cholesterol was elevated by19.6% and total cholesterol, total triglyceride, low density lipoproteincholesterol and Atherosclerosis Index were reduced by 23%, 36.5%, 28.5%and 34.2%, respectively. It was also observed that the higher theabnormality of the lipid and lipoprotein serum level, the more dramaticthe modulation of lipid and lipoprotein levels can be achieved byXuezhikang therapy. Xuezhikang can also reduce apolipoprotein B level,blood sedimentation and blood sedimentation K-value.

[0137] Overall, the results show that red rice was as a safe, effectiveagent for modulating serum lipid and lipoprotein levels. Red rice canalso be used as a therapeutic agent for coronary artery disease andcerebrovascular disease caused by hyperlipidemia and/or athyrosisbecause red rice not only significantly reduced plasma TC, TG andAtherosclerosis Index, but also markedly raised plasma apolipoprotein inas a-I level.

EXAMPLE 4 Human Clinical Trial II

[0138] In this clinical trial, 84 patients with hyperlipidemia, and 56patients who were also diagnosed with atherosclerosis were divided intotwo treatment groups: a group treated with Xuezhikang capsules and acontrol group treated with Jiaogulan tablets.

[0139] All patients were diagnosed as hyperlipidemic following thecriteria set forth in “Clinical trial management: hyperlipidemiatreatment using new Chinese materia medica” released by the Ministry ofHealth of China. After dietary advice for two to four weeks, bloodsamples were collected from patients with abnormal lipid and lipoproteintwice, two weeks prior to the trial. Only patients who met the followingcriteria were enrolled in the trial: total serum cholesterol (TC)>230mg/dl (5.95 mmol/L) and triglyceride (TG)>200 mg/dl (>2.26 mmol/L). Highdensity lipoprotein cholesterol (HDL-C) was also considered as areference: male<40 mg/dl (1.04 mmol/L); female<45 mg/dl (1.16 mmol/L).

[0140] The symptoms included: limb tightness; asthenic breathing; painand oppressed feeling in chest; loss of appetite; distention andswelling of gastric region; whitish or purple dots on tongue; thethick-white or thick-slimy fur on tongue; taut-slippery orhesitant-weak-pulse.

[0141] The severity of the symptoms as recognized by traditional ChineseMedicine were scored as follows: Asthenic breathing 0 none (−) noasthenic breathing 2 light (+) having asthenic breathing with physicalactivity 3 moderate (++) having medium asthenic breathing with physicalactivity 4 severe (+++) having asthenic breathing at rest Limb tightness0 none (−) no limb tightness 2 light (+) having limb tightnessoccasionally 3 moderate (++) having medium limb tightness very often 4severe (+++) having severe limb tightness Chest tightness and pain 0none (−) no chest tightness and pain 2 light (+) having chest tightnessand pain occasionally 3 moderate (++) having medium chest tightness andpain very often 4 severe (+++) having severe chest tightness and pain atrest Loss of appetite 0 none (−) having normal appetite 2 light (+)losing appetite by ¼-⅓ 3 moderate (++) losing appetite by ⅓-½ 4 severe(+++) losing appetite more than ½ Abdominal distention and swelling 0none (−) no this sign 2 light (+) having the sign occasionally 3moderate (++) having this sign very often 4 severe (+++) having severeabdominal distention and swelling Picture of the tongue 0 normal (−) 1abnormal (+) Pulse condition 0 normal (−) 1 abnormal (+)

[0142] Patients diagnosed by traditional Chinese medicine according tothe above symptoms, and patients with primary hyperlipidemia wereenrolled.

[0143] The criteria for exclusion of patients were as follows:

[0144] a. myocardial infarction, cerebrovascular disease, severe woundor major surgery during last half year;

[0145] b. nephritic syndrome, hypothyroidism, acute and/or chronichepatobiliary disorder, diabetes, gout;

[0146] c. familial hypercholesterolemia(monogenic-hypercho-lesterolemia);

[0147] d. secondary hyperlipidemia caused by other medication, forinstance: phenothiazine, beta-adrenergic blocking agents,corticosteroid, oral contraceptive;

[0148] e. patients who used other lipid modulators during the last fourweeks and patients using heparin or were on thyroidization;

[0149] f. pregnant and breast-feeding women;

[0150] g. patients with disorder of the other organs; and

[0151] h. hylaxis syndrome, and psychosis.

[0152] The total number of patients enrolled was 116. There were 84patients in the treated group and 32 patients in the control group. Nodifference of distribution in age, sex and course of disease were foundbetween the two groups.

[0153] A randomized single-blind trial was conducted with two groups.The treated group (84 cases) took two Xuezhikang capsules (i.e., a redrice product of the present invention) twice a day. The control group(32 cases) took three Jiaogulan tablets (ShanXi factory of Chinesemateria medica, lot number: 940730) twice a day. The course of treatmentwas eight weeks.

[0154] The measurements of serum lipid and lipoprotein levels and otherscoring were performed prior to the therapy, and at four weeks and ateight weeks after therapy. The safety tests were conducted before andafter therapy. A fasting venous blood sample was collected; patientswere not allowed to consume alcohol or food with a high fat content inthe last meal.

[0155] The following safety tests were conducted: blood and ureanitrogen (BUN); creatinine; serum glutamic pyruvic transminase (SGPT,ALT); serum glucose; and creatinine kinase (CK). Total serum cholesterol(TC), total serum triglyceride (TG) and high density lipoproteincholesterol levels were measured to determine efficacy. Other relevantclinical sign such as weight, high blood pressure, heart beat andrhythm, and hepatospleno-palpation were recorded.

[0156] Efficacy was evaluated according to the criteria set forth in“Clinical trial management: hyperlipidemia treatment using new Chinesemateria medica” released by the Ministry of Health of China as follows:

[0157] 1. Cure: All symptoms are eliminated or the total symptom scorereduced by more than 90%; and every laboratory tested parameters reachednormal levels.

[0158] 2. Effective: Symptoms are significantly relieved, i.e., symptomscore reduced by 70%-89%. Serum lipid and lipoprotein do not reachnormal level but was improved in one of the following respects: 1)reducing TC≧20%; 2) reducing TG≧40%; 3) reducing (TC−HDL-C)−/HDL-C≧20%;4) increasing HDL-C>10 mg/dl.

[0159] 3. Improvement: Symptoms are relieved, i.e., symptom scorereduced by 30%-69%. Serum lipid and lipoprotein did not reach normallevels but were improved in one of the following respects: 1) reducingTC at 10%-20%; 2) reducing TG≧20% but <40%; 3) reducing(TC−HDL-C)/HDL-C≧10% but <20%; 4) increasing HDL-C>4 mg/dl (0.14 mmmol/L).

[0160] 4. Inefficacy: Symptom score was reduced by less than 30% andlaboratory test parameters did not meet the criteria of effectiveness.

[0161] All the data were subjected to statistical analysis. Thestudent's t test, Chi-square test for counting data, and Ridit assaywere used when appropriate.

[0162] Table 5 shows a comparison of the overall efficacy score of thetwo groups. TABLE 5 General Efficacy Total Case Group Number CureEffective Improvement Inefficacy Treated 84 39 25 13 7 Control 32 3 6 419

[0163] The total efficacy score in the treated group was much higherthan that of the control group (X²−22.95, P<0.01).

[0164] Table 6 shows a comparison of efficacies as defined by thestandards of traditional Chinese medicine. TABLE 6 Comparison ofefficacy Red Rice Control Symptoms Before After Vanish % Before AfterVanish % Statistics Asthenic 35 15 57.1 12 6 50 >0.05 breathing Limbstight 37 16 56.8 13 7 46.2 >0.05 Oppressed 37 16 56.8 9 5 44.4 * feelingin chest Chest pain 8 1 87.5 2 1 50 * Loss of 11 4 63.6 3 2 33.3 *appetite Distension & 31 11 64.5 7 6 14.3 * swelling stomach Pale tongue54 34 37 17 12 31.3 >0.05 Purple dot 9 4 55.6 3 4 0 * on tongue Thick-21 16 23.8 11 6 45.5 >0.05 whitish fur Thick-slimy 11 8 27.3 5 4 0 furSlippery & 28 22 21.4 13 8 33.3 * string-like pulse Weak-thread 24 1633.3 8 3 62.6 * pulse Slippery- 28 16 42.9 9 8 11.1 * fine hpulse

[0165] The percentage of patients who reported elimination of thesymptoms as diagnosed by traditional Chinese medicine in the treatedgroup was much higher than that in the control group (p<0.05),especially in the aspect of pain and oppressed feeling in chest, loosingappetite, distention and swelling on gastric region as well as purplishdots on the tongue.

[0166] The change in serum total cholesterol level is shown in Table 7.TABLE 7 Change in serum total cholesterol level Abnormal ReductionReduction Reduction Group Case No. Cure >20% 10-20% <10% Treated 76 53 95 9 Control 28 4 0 2 22

[0167] The scores for curing or reducing total serum cholesterol levelin the treated group were greater than that in the control group.

[0168] The change in serum total triglyceride level is shown in Table 8.TABLE 8 Change in serum total triglyceride level Abnormal ReductionReduction Reduction Group Case No. Cure >20% 10-20% <10% Treated 35 20 25 9 Control 13 6 2 1 4

[0169] No significant difference in the scores for curing or reducingthe serum total triglyceride level was observed between the two groups.

[0170] The change in high density lipoprotein-cholesterol level is shownin Table 9. TABLE 9 Change in HDL-C levels. Abnormal (>4) Cure ReductionReduction Reduction Group Case No. >4 >20% 10-20% <10% Treated 24 11 1 39 Control 10 3 0 1 6

[0171] A similar efficacy for normalizing or increasing HDL-C level wasfound in both groups.

[0172] Table 10 shows the changes in Atherosclerotic Index which is theratio of (TC−HDL-C)/HDL-C. TABLE 10 Change of (TC-HDL-C)/HDL-C Abnormal(>4) Cure Reduction Reduction Reduction Group Case NO. >4 >20% 10-20%<10% Treated 56 44 6 3 3 Control 14 3 1 2 8

[0173] The data in Table 10 indicates that the both theXuezhikang-treated and control groups improved HDL-C serum levelsmarkedly. The effectiveness of Xuezhikang was found to be superior tothat of the control.

[0174] Table 11 shows the effect of Xuezhikang on regulating serum lipidand lipoprotein levels (X+/−S). TABLE 11 Regulating Effect of Xuezhikangon serum lipid and lipoprotein (X ± S) Case Case Case HDL-c Case GroupTime Point No. TC (mg/dl) No. TG (mg/dl) No. (mg/dl) No. LDL-c (mg/dl)Treated Baseline 76 273.9 ± 34.1 35 304.1 ± 86.8 24 35.3 ± 4.8 84174.7 + 48.1 Group 4 Weeks 253.9 ± 35.9 270.3 ± 121.2* 39.5 ± 9**141.4 + 477.6*** difference 35*** 33.8 4.2# 33.3### ↓13.87% ↓11.11%↑11.9% ↓19.1% 8 Weeks 216.7 ± 33.7 206.5 ± 72*** 42.1 ± 7.6** 126.8 +39.6*** difference 57.3*** 97.6 6.8 49.9### ↓20.91% ↓32.09% ↑19.6%↓27.4% Control Baseline 28 265.4 ± 25 13 297.1 ± 72.1 10 35.3 ± 3.3 32164.3 + 35.6 Group 4 Weeks 272.6 ± 33.3 304.8 ± 0.141 35.3 ± 3.7 171.6 +42.3 difference 7.2 7.7 0.02 7.3 ↑2.7 ↑2.6% ↓0.06% ↑4.44% 8 Weeks 265 ±35.8 226.7 ± 88.1 38.3 ± 7.3 168 + 45.5 difference 0.5 70.3 2.98 4.11↓0.2% ↓23.67% ↑8.43% ↑2.5%

[0175] Table 11 indicates that Xuezhikang improved TC, TG,(TC−HDL-C)/HDL-C, HDL-C serum levels markedly, but control group onlyimproved TG significantly. Xuezhikang therapy was found to regulate TC,LDL-C more effectively than the control.

[0176] Table 12 shows the efficacy of Xuezhikang therapy on differentbaseline of serum lipid and lipoprotein. TABLE 12 Efficacy comparison ofXuezhikang therapy on different baseline of serum lipid and lipoprotein.TC (mg/g) TG (mg/g) HDL-C (mg/dl) Parameter <230 230-300 >300 230230-300 >300 >45 35-45 <35 Case No. 8 60 16 49 20 15 55 17 12 Meanbaseline 192.01 261.8 327.1 134.3 247.6 327.3 56.4 40.1 5.4 (mean) Mean(4 weeks) 174.14 226.14 272.56 134.84 202.64 360.59 55.03 43.41 38.82 %Changes ↓9.31 ↓12.621 ↓17.62 ↓4.05 ↓16.2 ↓6.87 ↓2.68 ↑7.49 ↑24.05 Mean(8 weeks) 156.08 208.21 ↓86.1 119.38 169.51 255.88 57.58 46.89 40.33 %Changes ↓18.72 ↓19.53 ↓24.93 ↓15.05 ↓29.9 ↓33.91 ↑1.85 ↑16.11 ↑28.89Comparison * ** ** ** ** **

[0177] The higher baseline of TC and TG in the serum, the more reductionis achieved after using Xuezhikang.

[0178] The results indicate that the score of cure, and the score ofefficacy were 46.4% (38/84), 29.8% (25/84), respectively, in theXuezhikang-treated (red rice treated) group and 9.4% (3/32), 18.8%(6/32) in the control group. Total efficacy ratio in the treated group(72%) was much higher than that in the control group (28.2%, P<0.001).There were significant differences between the two groups in terms ofimproving TC, LDL-C and (TC−HDL-C)/HDL-C.

[0179] No significant clinically meaningful change in the followingparameters was observed during and after therapy: serum glutamic pyruvictransaminase (SGPT); blood and urea nitrogen (BUN); creatinine; serumglucose; cardioelectrogram; and routine examination of urine and blood.Three cases reported an increase in creatinine kinase (CK) (252, 260,466 IU/L versus normal standard at 200 IU/L) in the treated group andone (256 IU/L) in the control group. No clinical symptoms were observedin any of these cases. The results show that a red rice product of thepresent invention is a safe and acceptable lipid-modulating agent.

Other Embodiments

[0180] It is to be understood that while the invention has beendescribed in conjunction with the detailed description of thereof, theforegoing description is intended to illustrate and not limit the scopeof the invention, which is defined by the scope of the appended claims.Other aspects, advantages, and modifications are within the scope of thefollowing claims.

What is claimed:
 1. A composition for treating elevated serumcholesterol and/or triglycerides, said composition comprising: a redrice product containing at least about 0.05% lovastatin by weight. 2.The composition of claim 1, wherein said red rice product is the productof a process comprising: culturing a lovastatin-producing Monascusstrain in a culture medium comprising rice; removing said culture mediumto provide a solid residue; and sterilizing said solid residue toprovide said red rice product.
 3. The composition of claim 2, whereinsaid Monascus strain comprises a stain selected from the groupconsisting of Monascus albidus Sato AS 3.570, AS 3.4440, CGMCC No. 0317;Monascus pilosus Sato AS 3.4444, AS 3.4633, AS 3.4646, AS 3.4647;Monascus pubigerus Sato AS 3.4445; Monascus ruber van Tieghem AS 3.549,CGMCC No. 0315, CGMCC No. 0316; Monascus paxii Lingelsheim AS 3.4453;Monascus fuliginosus Sato AS 3.569, AS 3.1098, AS 3.2091, AS 3.2093, AS3.2134, IFFI 05035, and Monascus purpureus Went CGMCC No.
 0272. 4. Thecomposition of claim 2, wherein said process further comprises:extracting said solid residue with ethanol.
 5. A method for producingimproved red rice, said method comprising: providing alovastatin-producing Monascus strain; culturing said Monascus strain ina culture medium comprising rice at a temperature of about 15° C. toabout 35° C. for a period of about 2 to about 20 days to provide crudered yeast rice; and drying said crude red rice to provide improved redrice.
 6. The method of claim 5, wherein said Monascus strain comprises astrain selected from the group consisting of Monascus albidus Sato AS3.570, AS 3.4440, CGMCC No. 0317; Monascus pilosus Sato AS 3.4444, AS3.4633, AS 3.4646, AS 3.4647; Monascus pubigerus Sato AS 3.4445;Monascus ruber van Tieghem AS 3.549, CGMCC No. 0315, CGMCC No. 0316;Monascus paxii Lingelsheim AS 3.4453; Monascus fuliginosus Sato AS3.569, AS 3.1098, AS 3.2091, AS 3.2093, AS 3.2134,IFFI 05035, andMonacus purpureus Went CGMCC No.
 0272. 7. The method of claim 5, whereinsaid culture medium further comprises: about 2% to about 6% sugar; about2% to about 7% additional carbon source selected from the groupconsisting of glycerine, malt, potato juice; about 0% to about 3%peptone; about 0% to about 3% thick beef juice; and about 2% to about 4%defoamer.
 8. The method of claim 5, wherein said strain is cultured atabout 30° C. to about 34° C. for about 2 to about 4 days, and is thencultured at about 20° C. to about 25° C. for at least about 4 days. 9.The method of claim 5, which further comprises: extracting said improvedred rice with an ethanol solution to provide an ethanol extract; anddrying said extract.
 10. The method of claim 9, wherein said ethanolsolution comprises about 75% to about 95% aqueous ethanol.
 11. AMonascus strain capable of producing about 0.5% lovastatin uponculturing.
 12. The Monacus purpureus Went strain of claim 11, whereinsaid strain is selected from the group consisting of Monascus albidusSato AS 3.570, AS 3.4440, CGMCC No. 0317; Monascus pilosus Sato AS3.4444, AS 3.4633, AS 3.4646, AS 3.4647; Monascus pubigerus Sato AS3.4445; Monascus ruber van Tieghem AS 3.549, CGMCC No. 0315, CGMCC No.0316; Monascus paxii Lingelsheim AS 3.4453; Monascus fuliginosus Sato AS3.569, AS 3.1098, AS 3.2091, AS 3.2093, AS 3.2134, IFFI 05035, andMonascus purpureus Went CGMCC No.
 0272. 13. A method of treating orpreventing cardiovascular disorders in a mammal which comprises:administering to a mammal an effective amount of a red rice fermentationproduct comprising 0.5% lovastatin.
 14. The method of claim 13 whereinsaid red rice fermentation product is produced from the fermentation ofat least one lovastatin-producing Monascus strain.
 15. The method ofclaim 13 wherein said cardiovascular disorder is selected from the groupconsisting of myocardial infarction, coronary heart disease,hypertension, hypotension, atherosclerosis and arteriosclerosis.
 16. Amethod of treating or preventing stroke, cerebral thrombosis whichcomprises administering an effective amount of a red rice fermentationproduct.
 17. A method of treating or preventing fatty liver conditionsin a human which comprises administering to a human an effective amountof a red rice fermentation product.
 18. A method of reducing serumcholesterol and triglyceride levels to normal levels in a human in needof such therapy which comprises administering an effective amount of ared rice fermentation product.
 19. A pharmaceutical or nutritionalformulation for treating elevated serum cholesterol and/or triglyceridelevels, said formulation comprising: an effective amount of a red riceproduct; and a pharmaceutically acceptable excipient.
 20. Theformulation of claim 19, wherein said formulation comprises about 10 mglovastatin.